WO2016199877A1 - Crystal of 5-hydroxy-4-(trifluoromethyl)pyrazolopyridine derivative - Google Patents

Abstract

A crystal of a compound represented by general formula (I) (wherein R¹ represents a hydrogen atom or a hydroxyl group, and R represents a 2-(trifluoromethyl)pyrimidin-5-yl group or a 5-(trifluoromethyl)pyrazin-2-yl group) or a pharmacologically acceptable salt thereof, which has excellent LCAT activating activity and is useful as an active ingredient for a therapeutic agent or prophylactic agent for arteriosclerosis, arteriosclerotic heart diseases, coronary heart diseases (including heart failure, myocardial infarction, angina pectoris, cardiac ischemia, cardiovascular disorders and angioplasty restenosis), cerebrovascular diseases (including cerebral apoplexy and cerebral infarction), peripheral vascular diseases (including diabetic vascular complications), dyslipidemia, low-HDL cholesterolemia or kidney diseases, especially as an active ingredient for an anti-arteriosclerotic agent.

Description

Crystal of 5-hydroxy-4- (trifluoromethyl) pyrazolopyridine derivative

The present invention relates to a specific crystal of a pyrazolopyridine derivative or a pharmacologically acceptable salt thereof having an excellent lecithin cholesterol acetyltransferase (hereinafter referred to as LCAT) activating action (preferably a reversible LCAT activating action). About.

In advanced civilized countries, cardiovascular diseases (for example, heart disease, cerebrovascular disease, kidney disease, etc.) caused by hypertension, dyslipidemia, diabetes, etc. are a major problem. Antihypertensive drugs, antilipidemia drugs, and antidiabetic drugs are used for the treatment of hypertension, dyslipidemia, and hyperglycemia, respectively. Clinically, α and β blockers, diuretics, calcium antagonists, ACE inhibitors, and A-II antagonists are used as antihypertensive agents, and HMG-CoA reductase inhibitors, Insulin, sulfonylureas, metformin, glitazones, DPP4 inhibitors, and the like are used as antidiabetics such as ion exchange resins, nicotinic acid derivatives, probucol, and fibrates. These drugs contribute to the regulation of blood pressure and blood lipid or blood glucose levels. However, mortality due to heart disease, cerebrovascular disease and kidney disease has not been greatly improved even by the use of these drugs, and development of better therapeutic agents for these diseases is desired.

A direct risk factor for cardiovascular disease is arteriosclerosis accompanied by thickening of the arterial wall, and the cause of the thickening is due to accumulation of oxidized low density lipoprotein (hereinafter referred to as LDL) cholesterol in macrophages in the arterial wall. It is the formation of plaque (Non-Patent Documents 1 and 2). This plaque inhibits blood flow and promotes thrombus formation.

The results of many epidemiological studies show that the serum lipoprotein concentration is associated with diseases such as dyslipidemia and arteriosclerosis (for example, Non-Patent Document 3). An increase in the concentration of LDL cholesterol in the blood and a decrease in the concentration of high-density lipoprotein (hereinafter referred to as HDL) cholesterol are both risk factors for coronary artery disease.

Peripheral tissue cholesterol is extracted by HDL and esterified by lecithin cholesterol acetyltransferase (hereinafter referred to as LCAT) on HDL to become cholesteryl ester. Increased LCAT activity promotes the withdrawal of cholesterol from macrophages (for example, Non-Patent Documents 4 and 5). Therefore, a drug that enhances LCAT activity is considered to be useful as a medicament for the treatment or prevention of diseases such as dyslipidemia and arteriosclerosis.

Examples of known drugs that enhance LCAT activity include peptide compounds (for example, Non-patent Document 6) and, as small molecules, for example, compounds described in Patent Document 1.

As compounds having a pyrazolopyridine skeleton, compounds described in Patent Documents 2 and 3 are known.

WO2008 / 002591 pamphlet WO2012 / 028243 pamphlet WO2013 / 187462 pamphlet

Currently known compounds having an LCAT activating action are not satisfactory in terms of safety and effectiveness, and LCAT activators having excellent safety and effectiveness have been desired.

The present inventors have conducted various synthetic studies aiming at obtaining new anti-arteriosclerotic drugs by having an excellent LCAT activation action and promoting cholesterol withdrawal directly from macrophages. As a result, the present inventors have found that a pyrazolopyridine derivative having a specific structure or a pharmacologically acceptable salt thereof has an excellent LCAT activation action, and completed the present invention.

The present invention provides a specific crystal of a pyrazolopyridine derivative or a pharmacologically acceptable salt thereof having an excellent LCAT activating action (preferably a reversible LCAT activating action) and a medicament containing them. .

That is, the present invention
(1) General formula (I)

[Wherein, R ¹ represents a hydrogen atom or a hydroxyl group, and R represents a 2- (trifluoromethyl) pyrimidin-5-yl group or a 5- (trifluoromethyl) pyrazin-2-yl group. Or a pharmacologically acceptable salt thereof,
(2) The crystal according to (1), wherein R ¹ is a hydrogen atom, and R is a 2- (trifluoromethyl) pyrimidin-5-yl group,
(3) The crystal according to (1), wherein R ¹ is a hydroxyl group and R is a 5- (trifluoromethyl) pyrazin-2-yl group,
(4) (+)-4,5-dihydroxy-4- (trifluoromethyl) -3- {1- [5- (trifluoromethyl) pyrazin-2-yl] piperidin-4-yl} -1,4 , 5,7-Tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one or (+)-cis-5-hydroxy-4- (trifluoromethyl) -3- {1- [2 -(Trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one or a hydrate thereof Crystals of the
(5) In the powder X-ray diffraction pattern obtained by irradiation with copper Kα rays, the interplanar spacing d is 8.86, 7.19, 6.67, 6.07, 5.51, 4.76, 4.61. (+)-4,5-dihydroxy-4- (trifluoromethyl) -3- {1- [5- (trifluoromethyl) showing major peaks at 3.94, 3.79 and 3.47 angstroms ) Pyrazin-2-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one hydrate crystals,
(6) In the powder X-ray diffraction pattern obtained by irradiation with copper Kα rays, the interplanar spacing d is 15.28, 7.70, 6.67, 6.17, 5.62, 5.01, 4.58. (+)-4,5-dihydroxy-4- (trifluoromethyl) -3- {1- [5- (trifluoromethyl) showing major peaks at 4.43, 3.77 and 3.30 angstroms ) Pyrazin-2-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one hydrate crystals,
(7) In the powder X-ray diffraction pattern obtained by irradiation with copper Kα rays, the surface spacing d is 14.97, 11.50, 9.84, 9.50, 7.12, 5.73, 5.48. (+)-4,5-dihydroxy-4- (trifluoromethyl) -3- {1- [5- (trifluoromethyl) showing major peaks at 4.95, 4.46 and 3.81 Angstroms ) Pyrazin-2-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one hydrate crystals,
(8) In the powder X-ray diffraction pattern obtained by irradiation with copper Kα rays, the interplanar spacing d is 9.28, 7.28, 6.16, 5.96, 4.99, 4.86, 4.73. , 4.53, 3.48 and 3.04 Angstroms, showing (+)-4,5-dihydroxy-4- (trifluoromethyl) -3- {1- [5- (trifluoromethyl) ) Pyrazin-2-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one hydrate crystals,
(9) In the powder X-ray diffraction pattern obtained by irradiation with copper Kα rays, the surface spacing d is 18.79, 6.90, 6.21, 5.68, 5.56, 4.79, 4.56. (+)-4,5-dihydroxy-4- (trifluoromethyl) -3- {1- [5- (trifluoromethyl) showing major peaks at 4.44, 3.76 and 3.44 angstroms ) Pyrazin-2-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one,
(10) In the powder X-ray diffraction pattern obtained by irradiation with copper Kα rays, the interplanar spacing d is 9.09, 7.94, 7.58, 5.31, 4.97, 4.68, 4.60. , 4.50, 3.86 and 3.50 Angstroms, (+)-cis-5-hydroxy-4- (trifluoromethyl) -3- {1- [2- (trifluoromethyl ) Pyrimidin-5-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one hydrate crystals,
(11) In the powder X-ray diffraction pattern obtained by irradiation with copper Kα rays, the interplanar spacing d is 23.99, 5.96, 5.25, 5.19, 5.07, 4.94, 4.68. (+)-Cis-5-hydroxy-4- (trifluoromethyl) -3- {1- [2- (trifluoromethyl), showing major peaks at 4.58, 4.52 and 4.40 angstroms ) Pyrimidin-5-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one hydrate crystals,
(12) In the powder X-ray diffraction pattern obtained by irradiation with copper Kα rays, the interplanar spacing d is 15.60, 9.26, 7.14, 5.21, 4.07, 3.88, 3.41. (+)-Cis-5-hydroxy-4- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) showing major peaks at 3.24, 3.11 and 2.70 angstroms ) Pyrimidin-5-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one,
(13) In the powder X-ray diffraction pattern obtained by irradiation of copper Kα rays, the spacing d is 9.88, 6.11, 5.58, 5.09, 5.01, 4.92, 4.35. (+)-Cis-5-hydroxy-4- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) showing major peaks at 3.83, 3.76 and 3.28 angstroms ) Pyrimidin-5-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one,
(14) In the powder X-ray diffraction pattern obtained by irradiation with copper Kα rays, the interplanar spacing d is 8.29, 5.92, 5.51, 5.30, 4.76, 4.50, 4.26. (+)-Cis-5-hydroxy-4- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) showing major peaks at 3.98, 3.81 and 3.66 Å ) Pyrimidin-5-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one,
(15) In the powder X-ray diffraction pattern obtained by irradiation with copper Kα rays, the interplanar spacing d is 12.00, 8.53, 6.10, 5.42, 4.56, 4.32, 3.83. (+)-Cis-5R-hydroxy-4R- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) showing major peaks at 3.60, 3.39 and 3.04 angstroms ) Pyrimidin-5-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one,
(16) In the powder X-ray diffraction pattern obtained by irradiation with copper Kα rays, the interplanar spacing d is 10.62, 8.93, 8.26, 5.32, 5.25, 4.96, 4.58. , 4.52, 4.44 and 3.84 angstroms, (+)-cis-5R-hydroxy-4R- (trifluoromethyl) -3- {1- [2- (trifluoromethyl ) Pyrimidin-5-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one hydrate crystals,
(17) A pharmaceutical composition comprising the crystal according to any one of (1) to (16) as an active ingredient,
(18) For treatment or prevention of arteriosclerosis, arteriosclerotic heart disease, coronary heart disease, cerebrovascular disease, peripheral vascular disease, dyslipidemia, low HDL cholesterolemia, or renal disease (17 ) The pharmaceutical composition according to
(19) The pharmaceutical composition according to (17) for the treatment or prevention of arteriosclerosis,
(20) The pharmaceutical composition according to (17) for the treatment or prevention of dyslipidemia,
(21) The pharmaceutical composition according to (17) for the treatment or prevention of a disease caused by an increase in the concentration of LDL cholesterol in the blood,
(22) The pharmaceutical composition according to (17) for treating or preventing a disease caused by a decrease in the concentration of HDL cholesterol in the blood,
(23) an LCAT activator comprising the crystal according to any one of (1) to (16) as an active ingredient,
(24) A reversible LCAT activator comprising the crystal according to any one of (1) to (16) as an active ingredient,
(25) An anti-arteriosclerotic agent containing the crystal according to any one of (1) to (16) as an active ingredient,
(26) A prophylactic or therapeutic agent for arteriosclerosis comprising the crystal according to any one of (1) to (16) as an active ingredient,
(27) An agent for lowering the concentration of LDL cholesterol in blood, comprising the crystal according to any one of (1) to (16) as an active ingredient,
(28) An agent for increasing the concentration of HDL cholesterol in blood, comprising the crystal according to any one of (1) to (16) as an active ingredient,
(29) A pharmaceutical composition comprising the crystal according to any one of (1) to (16) and a pharmacologically acceptable carrier,
(30) Use of the crystal according to any one of (1) to (16) for the manufacture of a pharmaceutical composition,
(31) Pharmaceutical composition for treatment or prevention of arteriosclerosis, arteriosclerotic heart disease, coronary heart disease, cerebrovascular disease, peripheral vascular disease, dyslipidemia, low HDL cholesterolemia, or renal disease Use of (30) for the manufacture of goods,
(32) The use according to (30) for the manufacture of a pharmaceutical composition for the treatment or prevention of arteriosclerosis,
(33) The use according to (30) for the manufacture of a pharmaceutical composition for the treatment or prevention of dyslipidemia,
(34) Use according to (30) for the manufacture of a pharmaceutical composition for the treatment or prevention of a disease caused by an increase in the concentration of LDL cholesterol in the blood,
(35) Use according to (30) for the manufacture of a pharmaceutical composition for the treatment or prevention of a disease caused by a decrease in the concentration of HDL cholesterol in the blood,
(36) A method for activating LCAT, comprising administering an effective amount of the crystal according to any one of (1) to (16) to a human,
(37) A method for treating or preventing a disease, comprising administering an effective amount of the crystal according to any one of (1) to (16) to a human,
(38) The disease is arteriosclerosis, arteriosclerotic heart disease, coronary heart disease, cerebrovascular disease, peripheral vascular disease, dyslipidemia, low HDL cholesterolemia, or renal disease. (37) The method described in
(39) The method according to (37), wherein the disease is arteriosclerosis,
(40) The method according to (37), wherein the disease is dyslipidemia,
(41) The method according to (37), wherein the disease is a disease caused by an increase in the concentration of LDL cholesterol in the blood,
(42) The method according to (37), wherein the disease is a disease caused by a decrease in the concentration of HDL cholesterol in the blood,
(43) The crystal according to any one of (1) to (16), for use in a method for treatment or prevention of a disease,
(44) The disease is arteriosclerosis, arteriosclerotic heart disease, coronary heart disease, cerebrovascular disease, peripheral vascular disease, dyslipidemia, low HDL cholesterolemia, or renal disease. (43) The crystals described in
(45) The crystal according to (43), wherein the disease is arteriosclerosis,
(46) The crystal according to (43), wherein the disease is dyslipidemia,
(47) The crystal according to (43), wherein the disease is a disease caused by an increase in the concentration of LDL cholesterol in the blood, and
(48) The crystal according to (43), wherein the disease is a disease caused by a decrease in the concentration of HDL cholesterol in the blood.

Hereinafter, the compound (I) of the present invention will be described.

The compound (I) of the present invention includes a compound represented by the formula (I) and a tautomer thereof (Ix)

Including both compounds represented by: In the present application, unless otherwise specified, for convenience, the compound (I) containing any tautomer is represented by the structural formula of the formula (I) and the corresponding chemical name. In addition, any isomer of the other tautomer (amide-imidic acid) of the compound (I) of the present invention is contained in the present compound (I). In the present application, for convenience, a compound containing any tautomer (I) is also represented by the structural formula represented by formula (I) and the corresponding chemical name.

Since the compound (I) of the present invention has a basic group, it can be converted into an acid addition salt with a pharmacologically acceptable acid. In the present invention, “pharmacologically acceptable salts” include, for example, hydrohalides such as hydrofluoric acid salts, hydrochlorides, hydrobromides, hydroiodides; nitrates, perchlorates. Inorganic salts such as sulfates and phosphates; lower alkane sulfonates such as methane sulfonate, trifluoromethane sulfonate and ethane sulfonate; aryl sulfones such as benzene sulfonate and p-toluene sulfonate Acid salts; organic acid salts such as acetic acid, malic acid, fumarate, succinate, citrate, tartrate, succinate, maleate; and amino acid salts such as ornithate, glutamate, aspartate Can be mentioned.

The compound (I) of the present invention or a pharmacologically acceptable salt thereof may absorb water and become a hydrate when left in the atmosphere, and such a hydrate is also included in the present invention. Is done.

The compound (I) of the present invention or a pharmacologically acceptable salt thereof may become a solvate when left in a solvent, and such a solvate is also encompassed in the present invention.

Compound (I) of the present invention has an optical isomer based on an asymmetric center in the molecule. Unless otherwise specified, in the compounds of the present invention, these isomers and mixtures of these isomers are all represented by a single formula, that is, the general formula (I). Accordingly, the present invention includes all of these isomers and mixtures of these isomers.

Compound (I) of the present invention may also contain an unnatural proportion of atomic isotopes at one or more of the atoms constituting the compound. Examples of atomic isotopes include deuterium ( ² H), tritium ( ³ H), iodine-125 ( ¹²⁵ I), carbon-14 ( ¹⁴ C), and the like. In addition, the compound may be radiolabeled with a radioisotope such as tritium ( ³ H), iodine-125 ( ¹²⁵ I), or carbon-14 ( ¹⁴ C). Radiolabeled compounds are useful as therapeutic or prophylactic agents, research reagents such as assay reagents, and diagnostic agents such as in vivo diagnostic imaging agents. All isotope variants of the compounds of the present invention, whether radioactive or not, are intended to be included within the scope of the present invention.

In the present invention, the salt of compound (I) or a hydrate or solvate thereof produces crystals (crystal polymorphs) having a plurality of different internal structures and physicochemical properties depending on the reaction conditions and crystallization conditions. Each of those crystals or mixtures thereof in any proportion are encompassed by the present invention. Moreover, although a crystalline solid and an amorphous (amorphous) solid may be mixed, any mixture thereof is included in the present invention. That is, the crystal having a specific crystal form of the present invention may contain a crystal having another crystal form or an amorphous solid, and the content of the specific crystal form is preferably 50% or more. More preferably, it is 80% or more, more preferably 90% or more, still more preferably 93% or more, particularly preferably 95% or more, and most preferably Is 97% or more.

In the present invention, a crystal indicates a solid whose internal structure is composed of regular repetitions of constituent atoms (or a group thereof) in a three-dimensional manner, and is distinguished from an amorphous solid that does not have such a regular internal structure. The Whether or not a certain solid is a crystal can be examined by a crystallographically well-known method (for example, powder X-ray crystal analysis, differential scanning calorimetry, etc.). For example, powder X-ray crystal analysis using X-rays obtained by irradiation with copper Kα rays is performed on a solid, and when a clear peak is observed in the X-ray diffraction pattern, the solid is determined to be a crystal. If no clear peak is observed, the solid is determined to be amorphous. If the peak can be read but the peak is not clear (eg, broad), the solid is determined to be a crystal with low crystallinity, and such low crystal with crystal Are included in the crystal.

In powder X-ray crystal analysis using copper Kα rays, the sample is usually irradiated with copper Kα rays (Kα1 and Kα2 rays are not separated). The X-ray diffraction diagram can be obtained by analyzing the diffraction derived from the Kα ray, or can be obtained by analyzing only the diffraction derived from the Kα1 ray extracted from the diffraction derived from the Kα ray. In the present invention, the powder X-ray diffraction diagram obtained by irradiation with Kα rays is an X-ray diffraction diagram obtained by analyzing diffraction derived from Kα rays, and X obtained by analyzing diffraction derived from Kα1 rays. It is an X-ray diffraction diagram including a line diffraction diagram, and preferably obtained by analyzing diffraction derived from the Kα1 line.

(+)-4,5-dihydroxy-4- (trifluoromethyl) -3- {1- [5- (trifluoromethyl) pyrazin-2-yl] piperidin-4-yl of the compound (I) of the present invention } -1,4,5,7-Tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one Crystals of hydrates are irradiated with copper Kα rays, for example, as shown in FIG. In the powder X-ray diffraction pattern obtained in the above, the interplanar spacing d is 8.86, 7.19, 6.67, 6.07, 5.51, 4.76, 4.61, 3.94, 3.79 and It may be a crystal showing a major peak at 3.47 angstroms. Here, the main peak is a peak having a relative intensity of 13 or more when the intensity of a peak having an interplanar spacing d of 4.76 angstroms is defined as 100. In the following powder X-ray diffraction diagrams of FIGS. 1 to 12, the vertical axis represents the diffraction intensity [count / second (cps)], and the horizontal axis represents the diffraction angle 2θ (degrees). Further, the surface interval d (angstrom) can be calculated as n = 1 in the equation 2dsin θ = nλ. In the above formula, the wavelength λ of the Kα line is 1.54 angstroms, and the wavelength λ of the Kα1 line is 1.541 angstroms. Since the position and relative intensity of the interplanar spacing d can vary somewhat depending on the measurement conditions and the like, even if the interplanar spacing d is slightly different, the crystal form is identical with reference to the entire spectrum pattern as appropriate. Sex should be certified.

(+)-4,5-dihydroxy-4- (trifluoromethyl) -3- {1- [5- (trifluoromethyl) pyrazin-2-yl] piperidin-4-yl of the compound (I) of the present invention } -1,4,5,7-Tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one Crystals of hydrates are irradiated with copper Kα rays, for example, as shown in FIG. In the powder X-ray diffraction pattern obtained in the above, the interplanar spacing d is 15.28, 7.70, 6.67, 6.17, 5.62, 5.01, 4.58, 4.43, 3.77 and It may be a crystal showing a major peak at 3.30 angstroms. Here, the main peak is a peak having a relative intensity of 20 or more when the intensity of a peak having an interplanar spacing d of 5.01 angstroms is defined as 100.

(+)-4,5-dihydroxy-4- (trifluoromethyl) -3- {1- [5- (trifluoromethyl) pyrazin-2-yl] piperidin-4-yl of the compound (I) of the present invention } -1,4,5,7-Tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one Crystals of hydrates are irradiated with copper Kα rays, for example, as shown in FIG. In the powder X-ray diffraction pattern obtained in the above, the interplanar spacing d is 14.97, 11.50, 9.84, 9.50, 7.12, 5.73, 5.48, 4.95, 4.46 and It may be a crystal showing a major peak at 3.81 Å. Here, the main peak is a peak having a relative intensity of 12 or more when the intensity of a peak having an interplanar spacing d of 14.97 angstroms is defined as 100.

(+)-4,5-dihydroxy-4- (trifluoromethyl) -3- {1- [5- (trifluoromethyl) pyrazin-2-yl] piperidin-4-yl of the compound (I) of the present invention } -1,4,5,7-Tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one Crystals of hydrates are irradiated with copper Kα rays, for example, as shown in FIG. In the powder X-ray diffraction pattern obtained in the above, the interplanar spacing d is 9.28, 7.28, 6.16, 5.96, 4.99, 4.86, 4.73, 4.53, 3.48 and It may be a crystal showing a major peak at 3.04 angstroms. Here, the main peak is a peak having a relative intensity of 11 or more when the intensity of a peak having an interplanar spacing d of 4.73 angstroms is defined as 100.

(+)-4,5-dihydroxy-4- (trifluoromethyl) -3- {1- [5- (trifluoromethyl) pyrazin-2-yl] piperidin-4-yl of the compound (I) of the present invention } -1,4,5,7-Tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one Anhydrous crystals, for example, are irradiated with copper Kα rays as shown in FIG. In the powder X-ray diffraction pattern obtained in the above, the interplanar spacing d is 18.79, 6.90, 6.21, 5.68, 5.56, 4.79, 4.56, 4.44, 3.76 and It may be a crystal showing a major peak at 3.44 angstroms. Here, the main peak is a peak having a relative intensity of 17 or more when the intensity of a peak having an interplanar spacing d of 4.79 angstroms is defined as 100.

(+)-Cis-5-hydroxy-4- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl of the compound (I) of the present invention } -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one Crystals of hydrates are irradiated with copper Kα rays, for example, as shown in FIG. In the powder X-ray diffraction pattern obtained in the above, the interplanar spacing d is 9.09, 7.94, 7.58, 5.31, 4.97, 4.68, 4.60, 4.50, 3.86 and It may be a crystal showing a major peak at 3.50 angstroms. Here, the main peak is a peak having a relative intensity of 20 or more when the intensity of a peak having an interplanar spacing d of 4.50 angstroms is defined as 100.

(+)-Cis-5-hydroxy-4- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl of the compound (I) of the present invention } -1,4,5,7-Tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one Crystals of hydrates are irradiated with copper Kα rays, for example, as shown in FIG. In the powder X-ray diffraction pattern obtained in the above, the interplanar spacing d is 23.99, 5.96, 5.25, 5.19, 5.07, 4.94, 4.68, 4.58, 4.52, and It may be a crystal showing a major peak at 4.40 angstroms. Here, the main peak is a peak having a relative intensity of 58 or more when the intensity of a peak having an interplanar spacing d of 4.40 angstroms is defined as 100.

(+)-Cis-5-hydroxy-4- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl of the compound (I) of the present invention } -1,4,5,7-Tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one Anhydrous crystals, for example, are irradiated with copper Kα rays as shown in FIG. In the powder X-ray diffraction pattern obtained in the above, the interplanar spacing d is 15.60, 9.26, 7.14, 5.21, 4.07, 3.88, 3.41, 3.24, 3.11, and It may be a crystal showing a major peak at 2.70 angstroms. Here, the main peak is a peak having a relative intensity of 4 or more when the intensity of a peak having an interplanar spacing d of 15.60 angstroms is defined as 100.

(+)-Cis-5-hydroxy-4- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl of the compound (I) of the present invention } -1,4,5,7-Tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one Anhydrous crystals, for example, are irradiated with copper Kα rays as shown in FIG. In the powder X-ray diffraction diagram obtained in (1), the interplanar spacing d is 9.88, 6.11, 5.58, 5.09, 5.01, 4.92, 4.35, 3.83, 3.76 and It may be a crystal showing a major peak at 3.28 angstroms. Here, the main peak is a peak having a relative intensity of 30 or more when the intensity of a peak having an interplanar spacing d of 4.35 angstroms is defined as 100.

(+)-Cis-5-hydroxy-4- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl of the compound (I) of the present invention } -1,4,5,7-Tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one Anhydrous crystals, for example, are irradiated with copper Kα rays as shown in FIG. In the powder X-ray diffraction pattern obtained in the above, the interplanar spacing d is 8.29, 5.92, 5.51, 5.30, 4.76, 4.50, 4.26, 3.98, 3.81 and It may be a crystal showing a major peak at 3.66 angstroms. Here, the main peak is a peak having a relative intensity of 23 or more when the intensity of a peak having an interplanar spacing d of 4.50 angstroms is defined as 100.

(+)-Cis-5R-hydroxy-4R- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl of the compound (I) of the present invention } -1,4,5,7-Tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one Anhydrous crystals, for example, are irradiated with copper Kα rays as shown in FIG. In the powder X-ray diffraction pattern obtained in the above, the interplanar spacing d is 12.00, 8.53, 6.10, 5.42, 4.56, 4.32, 3.83, 3.60, 3.39 and It may be a crystal showing a major peak at 3.04 angstroms. Here, the main peak is a peak having a relative intensity of 11 or more when the intensity of a peak having an interplanar spacing d of 5.42 angstroms is defined as 100.

(+)-Cis-5R-hydroxy-4R- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl of the compound (I) of the present invention } -1,4,5,7-Tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one Crystals of hydrate are, for example, irradiated with copper Kα rays as shown in FIG. In the powder X-ray diffraction pattern obtained in the above, the interplanar spacing d is 10.62, 8.93, 8.26, 5.32, 5.25, 4.96, 4.58, 4.52, 4.44 and It may be a crystal showing a major peak at 3.84 angstroms. Here, the main peak is a peak having a relative intensity of 28 or more when the intensity of a peak having an interplanar spacing d of 10.62 angstroms is defined as 100.

The crystal of the compound represented by the general formula (I) of the present invention or a pharmacologically acceptable salt thereof has an excellent LCAT activating action, and it is arteriosclerosis, arteriosclerotic heart disease, coronary heart disease. (Including heart failure, myocardial infarction, angina pectoris, cardiac ischemia, cardiovascular disorder and angiogenic restenosis), cerebrovascular disease (including stroke and cerebral infarction), peripheral vascular disease (including diabetic vascular complications), It is useful as an active ingredient of a therapeutic or prophylactic agent for dyslipidemia, low HDL cholesterolemia, or renal disease, particularly an anti-atherosclerotic agent.

(+)-4,5-dihydroxy-4- (trifluoromethyl) -3- {1- [5- (trifluoromethyl) pyrazin-2-yl] piperidin-4-yl} obtained in Example 1 X-ray powder diffraction pattern of -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one hydrate. (+)-4,5-Dihydroxy-4- (trifluoromethyl) -3- {1- [5- (trifluoromethyl) pyrazin-2-yl] piperidin-4-yl} obtained in Example 2 X-ray powder diffraction pattern of -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one hydrate. (+)-4,5-dihydroxy-4- (trifluoromethyl) -3- {1- [5- (trifluoromethyl) pyrazin-2-yl] piperidin-4-yl} obtained in Example 3 X-ray powder diffraction pattern of -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one hydrate. (+)-4,5-dihydroxy-4- (trifluoromethyl) -3- {1- [5- (trifluoromethyl) pyrazin-2-yl] piperidin-4-yl} obtained in Example 4 X-ray powder diffraction pattern of -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one hydrate. (+)-4,5-dihydroxy-4- (trifluoromethyl) -3- {1- [5- (trifluoromethyl) pyrazin-2-yl] piperidin-4-yl} obtained in Example 5 X-ray powder diffraction pattern of -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one. (+)-Cis-5-Hydroxy-4- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl} obtained in Example 6 X-ray powder diffraction pattern of -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one hydrate. (+)-Cis-5-Hydroxy-4- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl} obtained in Example 7 X-ray powder diffraction pattern of -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one hydrate. (+)-Cis-5-Hydroxy-4- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl} obtained in Example 8 X-ray powder diffraction pattern of -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one. (+)-Cis-5-Hydroxy-4- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl} obtained in Example 9 X-ray powder diffraction pattern of -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one. (+)-Cis-5-hydroxy-4- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl} obtained in Example 10 X-ray powder diffraction pattern of -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one. (+)-Cis-5R-hydroxy-4R- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl} obtained in Example 11 X-ray powder diffraction pattern of -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one. (+)-Cis-5R-hydroxy-4R- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl} obtained in Example 12 X-ray powder diffraction pattern of -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one hydrate. Dose response curves for determining the 50% effective concentration of LCAT activation in Test Example 1 and 2 of the present invention _{(EC 50).}

Compound (I) of the present invention can be produced by the method described in the examples.

In the production process of compound (I) of the present invention, the product of each step is used as a free compound or a salt thereof, after completion of the reaction, if necessary, in a conventional manner, for example, (1) a method of concentrating the reaction solution as it is. (2) A method of removing insoluble matters such as a catalyst by filtration, and concentrating the filtrate. (3) Water and a solvent immiscible with water (for example, dichloroethane, diethyl ether, ethyl acetate, toluene, etc.) are added to the reaction solution. In addition, it can be isolated from the reaction mixture by a method for extracting the product, (4) a method for filtering the crystallized or precipitated product, and the like. The isolated product can be purified by a conventional method such as recrystallization, reprecipitation, various chromatographies and the like, if necessary. Alternatively, the product of each step can be used in the next step without isolation or purification.

Compound (I) of the present invention is isolated and purified as a free compound, a pharmacologically acceptable salt, hydrate, or solvate thereof. The pharmacologically acceptable salt of the compound (I) of the present invention can be produced by subjecting it to a conventional salt formation reaction. Isolation and purification are carried out by applying ordinary chemical operations such as extraction, concentration, distillation, crystallization, filtration, recrystallization, or various chromatography.

Various isomers can be separated by utilizing differences in physicochemical properties between isomers. For example, a racemic mixture can be converted to an optically pure isomer, such as by fractional crystallization leading to a diastereomeric salt with an optically active base or acid, or chromatography using a chiral column. Further, the diastereo mixture can be separated by fractional crystallization or various chromatographies. An optically active compound can also be produced by using an appropriate optically active raw material.

Examples of the administration form of the compound having the general formula (I) of the present invention or a pharmacologically acceptable salt thereof include oral administration by tablet, granule, powder, capsule or syrup, or injection or suppository. Parenteral administration, and the like, and can be administered systemically or locally.

Examples of the oral pharmaceutical form of the compound having the general formula (I) of the present invention or a pharmacologically acceptable salt thereof include tablets, pills, granules, powders, capsules, solutions, suspensions, emulsions, Examples include syrups and elixirs. Examples of pharmaceutical forms for parenteral use include injections, ointments, gels, creams, patches, sprays, inhalants, sprays, eye drops, and suppositories. These forms of pharmaceuticals are pharmaceutically acceptable, such as excipients, binders, diluents, stabilizers, preservatives, colorants, solubilizers, suspending agents, buffering agents, or wetting agents. The additive can be prepared according to a conventional method using additives appropriately selected as necessary.

The dosage of the compound having the general formula (I) of the present invention or a pharmacologically acceptable salt thereof is as follows: symptoms, body weight, age, administration method of the administered person (warm-blooded animal, eg, human) Varies depending on etc. For example, in the case of oral administration, the lower limit is 0.01 mg / kg body weight (preferably 0.03 mg / kg body weight) and the upper limit is 300 mg / kg body weight (preferably 100 mg / kg body weight). It is desirable to administer one to several times a day depending on the symptoms. In the case of intravenous administration, the lower limit is 0.01 mg / kg body weight (preferably 0.03 mg / kg body weight) and the upper limit is 300 mg / kg body weight (preferably 100 mg / kg body weight). Is preferably administered one to several times per day depending on the symptoms.

Hereinafter, the present invention will be described in more detail with reference to Examples, Test Examples and Formulation Examples, but the scope of the present invention is not limited thereto. In the following examples, hexane represents n-hexane, THF represents tetrahydrofuran, IPA represents 2-propanol, DMF represents N, N'-dimethylformamide, and DMSO represents dimethyl sulfoxide.

Example 1 (+)-4,5-dihydroxy-4- (trifluoromethyl) -3- {1- [5- (trifluoromethyl) pyrazin-2-yl] piperidin-4-yl} -1 , 4,5,7-Tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one hydrate (1) 4- [5-amino-1- (diphenylmethyl) -1H-pyrazole-3- Yl] piperidine-1-carboxylate

To a solution of tert-butyl 4- (cyanoacetyl) piperidine-1-carboxylate (compound described in WO 2004/14910 pamphlet, 7.1 g, 28 mmol) in ethanol (71 mL) was added diphenylmethylhydrazine hydrochloride (8.57 g). 36.5 mmol), and stirred at 50 ° C. for 1 hour. The reaction solution was concentrated under reduced pressure, and saturated aqueous sodium hydrogen carbonate solution and ethyl acetate were added to the resulting residue for liquid separation, the organic layer was dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. . The obtained residue was purified by silica gel column chromatography [eluent: hexane / ethyl acetate = 95 / 5-40 / 60 (gradient)] to obtain the title compound (7.43 g, yield: 59%). .

¹ H-NMR (400Hz, CDCl ₃ ) δ: 7.37-7.19 (10H, m), 6.66 (1H, s), 5.40 (1H, s), 4.11 (1H, brs), 3.23-3.20 (1H, m) , 2.82-2.65 (3H, m), 1.89-1.86 (2H, m), 1.61-1.52 (4H, m), 1.46 (9H, s).

(2) 4- [4,5-dihydroxy-6-oxo-4- (trifluoromethyl) -4,5,6,7-tetrahydro-1H-pyrazolo [3,4-b] pyridin-3-yl] Piperidine-1-carboxylate tert-butyl

To a suspension of sodium hydride (63% oil dispersion, 5.127 g, 134.6 mmol) in toluene (80 mL) at room temperature, ethyl 2-triethylsilyloxyacetate (reference J. Org. Chem., 2008, No. 1). 73, 6268-6278, 18.37 g, 84.13 mmol), and a solution of ethanol (0.1474 mL, 2.524 mmol) in toluene (40 mL) followed by ethyl trifluoroacetate. A toluene (20 mL) solution of (15.07 mL, 126.2 mmol) was added and stirred for 5 minutes, and then stirred at 80 ° C. for 30 minutes. Saturated aqueous ammonium chloride solution was added to the reaction mixture under ice-cooling, and the mixture was extracted with ethyl acetate. The resulting organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. An oily crude product (23.6 g) was obtained.

An oily crude product (23.6 g) obtained by the above operation and 4- [5-amino-1- (diphenylmethyl) -1H-pyrazol-3-yl] piperidine produced in (1) A mixed solution of tert-butyl-1-carboxylate (10.83 g, 25.04 mmol) in ethanol (150 mL) and acetic acid (50 mL) was stirred with heating under reflux for 4 hours. The solvent of the reaction solution was distilled off under reduced pressure, ethyl acetate was added to the resulting residue, washed successively with saturated aqueous sodium hydrogen carbonate solution and saturated brine, dried over anhydrous sodium sulfate, and dried under reduced pressure. The solvent was distilled off. Ethyl acetate and hexane were added to the resulting residue, and the resulting precipitate was collected by filtration to obtain a solid. Further, the solvent of the filtrate was distilled off under reduced pressure, and the obtained residue was purified by silica gel column chromatography [eluent: hexane / ethyl acetate = 90 / 10-50 / 50 (gradient)]. In combination with the previously obtained solid, a synthetic intermediate was obtained.

Triethylsilane (10.7 mL, 67.4 mmol) and trifluoroacetic acid (90 mL, 1176 mmol) were added to a suspension of the synthetic intermediate obtained in the above operation in dichloromethane (300 mL), and the mixture was stirred at room temperature for 2 hours. Stir. The solvent of the reaction solution was distilled off under reduced pressure, diethyl ether and hexane were added to the resulting residue, and the mixture was stirred for 30 minutes. The resulting precipitate was collected by filtration to obtain a colorless solid.

A colorless solid ethyl acetate (120 mL) and THF (40 mL) mixed suspension obtained by the above operation were mixed with di-t-butyl dicarbonate (5.53 g, 25.4 mmol) at room temperature, and , A solution of triethylamine (4.69 mL, 33.8 mmol) in ethyl acetate (30 mL) was added, stirred at room temperature for 3 hours, and allowed to stand at room temperature overnight. The reaction mixture was washed successively with saturated aqueous sodium hydrogen carbonate solution and saturated brine, dried over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure, and the obtained residue was purified by silica gel column chromatography [eluent: hexane. / Ethyl acetate = 50: 50-0: 100 (gradient)] to obtain the title compound (3.09 g, yield: 44%).

¹ H-NMR (400MHz, DMSO-d ₆ ) δ: 12.24 (1H, s), 10.55 (1H, s), 6.76 (1H, s), 5.65 (1H, d, J = 4Hz), 4.33 (1H, brs), 4.13-3.99 (2H, m), 3.20-3.09 (1H, m), 2.84-2.59 (2H, m), 1.84-1.76 (1H, m), 1.66-1.46 (3H, m), 1.42 ( 9H, s).

(3) 4- [4,5-dihydroxy-6-oxo-4- (trifluoromethyl) -4,5,6,7-tetrahydro-1H-pyrazolo [3,4-b] pyridin-3-yl] Optically active form of tert-butyl piperidine-1-carboxylate

4- [4,5-Dihydroxy-6-oxo-4- (trifluoromethyl) -4,5,6,7-tetrahydro-1H-pyrazolo [3,4-b] pyridine prepared in (2) A mixed solution of tert-butyl-3-yl] piperidine-1-carboxylate (0.79 g, 1.9 mmol) in ethyl acetate and methanol was adsorbed onto silica gel, and the solvent was distilled off under reduced pressure. The obtained powder was purified by flash LC [column: Chiralflash IA (30 mm id × 100 mm); manufactured by Daicel, elution solvent: hexane / IPA = 90/10, flow rate: 12 mL / min] to give the title compound (0 .34 g, yield: 43%, optically active substance).

Optical purity was measured using HPLC [column: Chiralpak IA (4.6 mm id x 250 mm); manufactured by Daicel, elution solvent: hexane / IPA = 80/20, flow rate 1.0 mL / min].

Optical purity 99% or more (retention time: 7.7 minutes).

(4) (+)-4,5-Dihydroxy-4- (trifluoromethyl) -3- {1- [5- (trifluoromethyl) pyrazin-2-yl] piperidin-4-yl} -1,4 , 5,7-Tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one

4- [4,5-Dihydroxy-6-oxo-4- (trifluoromethyl) -4,5,6,7-tetrahydro-1H-pyrazolo [3,4-b] pyridine prepared in (3) Trifluoroacetic acid (2 mL) was added to a suspension of an optically active form of tert-butyl (-3-yl) piperidine-1-carboxylate (0.34 g, 0.81 mmol) in dichloromethane (6 mL) at room temperature. Stir for 2 hours. The solvent of the reaction solution was distilled off under reduced pressure, diethyl ether and hexane were added to the resulting residue to solidify, the solvent was removed by decantation, and the resulting solid was dried under reduced pressure to obtain a synthetic intermediate. It was.

To a DMSO (5 mL) solution of the synthetic intermediate obtained by the above operation, at room temperature, 2-chloro-5- (trifluoromethyl) pyrazine (0.15 mL, 1.2 mmol) and N, N— Diisopropylethylamine (0.41 mL, 2.4 mmol) was added, and the mixture was stirred for 1 hour and then left overnight. Ethyl acetate was added to the reaction mixture, washed successively with water and saturated brine, dried over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography [eluent: hexane. / Ethyl acetate = 50 / 50-0 / 1000 (gradient)] to give the title compound (0.31 g, yield: 82%, optically active substance).

Optical purity was measured using HPLC [column: Chiralpak IA (4.6 mm id x 250 mm); manufactured by Daicel, elution solvent: hexane / IPA = 60/40, flow rate 1.0 mL / min].

Optical purity 99% or more (retention time: 6.7 minutes);
¹ H-NMR (400MHz, DMSO-d ₆ ) δ: 12.21 (1H, s), 10.57 (1H, s), 8.51 (1H, s), 8.50 (1H, s), 6.81 (1H, s), 5.68 (1H, d, J = 4Hz), 4.69-4.56 (2H, m), 4.37-4.31 (1H, m), 3.46-3.34 (1H, m), 3.11-2.97 (2H, m), 1.98-1.88 ( 1H, m), 1.83-1.58 (3H, m);
MS (ESI) m / z: 467 (M + H) ⁺ ;
[α] _D ²⁵ = + 3.9 ° (DMF, c = 0.924).

(5) (+)-4,5-Dihydroxy-4- (trifluoromethyl) -3- {1- [5- (trifluoromethyl) pyrazin-2-yl] piperidin-4-yl} -1,4 , 5,7-Tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one hydrate (+)-4,5-dihydroxy-4- (trifluoromethyl) prepared in (4) -3- {1- [5- (trifluoromethyl) pyrazin-2-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridine-6 Toluene (395 μl) was added to -one (19.73 mg) at room temperature. Thereafter, the mixture was stirred at 40 ° C. for about 20 hours, and then stirred at room temperature for about 0.5 hour. The solid was collected by filtration and dried overnight at room temperature to obtain the title compound (15.71 mg). Recovery rate 78%.

Calculated as elemental analysis value C ₁₇ H ₁₆ F ₆ N ₆ O ₃ .0.5H ₂ O: C, 42.95; H, 3.60; F, 23.98; N, 17.68.
Found: C, 42.97; H, 3.58; F, 23.79; N, 17.21.

1 shows a diffraction pattern of powder X-ray diffraction (CuKα, λ = 1.54 Å, scanning speed = 20 ° / min) with a relative intensity of 13 or more when the maximum peak intensity is 100 in FIG.

(Table 1)
――――――――――――――――――――――――――――――
Peak number 2θ d value Relative intensity ――――――――――――――――――――――――――――――
1 9.98 8.86 40
2 12.30 7.19 21
3 13.26 6.67 17
4 14.58 6.07 64
5 16.08 5.51 13
6 18.64 4.76 100
7 19.24 4.61 22
8 22.56 3.94 21
9 23.48 3.79 14
10 25.62 3.47 15
――――――――――――――――――――――――――――――

Example 2 (+)-4,5-Dihydroxy-4- (trifluoromethyl) -3- {1- [5- (trifluoromethyl) pyrazin-2-yl] piperidin-4-yl} -1 , 4,5,7-Tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one hydrate (+)-4,5-dihydroxy-4-prepared in Example 1 (4) (Trifluoromethyl) -3- {1- [5- (trifluoromethyl) pyrazin-2-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4 b] 1,4-Dioxane (2 ml) was added to pyridin-6-one (21.21 mg) at room temperature and completely dissolved. Then, after freezing in a freezer at −80 ° C., the solvent was removed while changing the temperature from −45 ° C. to 25 ° C. with a freeze dryer. Next, the glass vial containing the freeze-dried specimen was opened and placed in a screw tube containing water. The screw tube was sealed and stored in a thermostatic bath at 25 ° C. for 3 days. Thereafter, the glass vial was taken out and air-dried overnight to obtain the title compound.

Calculated as elemental analysis value C ₁₇ H ₁₆ F ₆ N ₆ O ₃ .1.8H ₂ O: C, 40.94; H, 3.96; F, 22.85; N, 16.85.
Found: C, 40.63; H, 3.83; F, 23.11; N, 16.57.

Table 2 shows the diffraction pattern of powder X-ray diffraction (CuKα, λ = 1.54 Å, scanning speed = 20 ° / min) and the peak with a relative intensity of 20 or more when the maximum peak intensity is 100 in FIG.

(Table 2)
――――――――――――――――――――――――――――――
Peak number 2θ d value Relative intensity ――――――――――――――――――――――――――――――
1 5.78 15.28 28
2 11.48 7.70 66
3 13.26 6.67 24
4 14.34 6.17 32
5 15.76 5.62 20
6 17.68 5.01 100
7 19.36 4.58 34
8 20.04 4.43 52
9 23.58 3.77 43
10 26.98 3.30 30
――――――――――――――――――――――――――――――

Example 3 (+)-4,5-Dihydroxy-4- (trifluoromethyl) -3- {1- [5- (trifluoromethyl) pyrazin-2-yl] piperidin-4-yl} -1 , 4,5,7-Tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one hydrate (+)-4,5-dihydroxy-4-prepared in Example 1 (4) (Trifluoromethyl) -3- {1- [5- (trifluoromethyl) pyrazin-2-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4 b] Nitromethane (200 μl) was added to pyridin-6-one (19.97 mg) at room temperature. Thereafter, the mixture was stirred at 10 ° C. for about 20 hours, and then stirred at room temperature for about 0.5 hour. The solid was collected by filtration and dried overnight at room temperature to obtain the title compound (12.78 mg). Recovery rate 62%.

Calculated as elemental analysis value C ₁₇ H ₁₆ F ₆ N ₆ O ₃ .1.0H ₂ O: C, 42.16; H, 3.75; F, 23.53; N, 17.35.
Found: C, 42.47; H, 3.46; F, 23.65; N, 17.39.

Table 3 shows the peak of relative intensity of 12 or more when the diffraction peak pattern of powder X-ray diffraction (CuKα, λ = 1.54 Å, scanning speed = 20 ° / min) in FIG.

(Table 3)
――――――――――――――――――――――――――――――
Peak number 2θ d value Relative intensity ――――――――――――――――――――――――――――――
1 5.90 14.97 100
2 7.68 11.50 22
3 8.98 9.84 63
4 9.30 9.50 57
5 12.42 7.12 14
6 15.46 5.73 72
7 16.16 5.48 15
8 17.90 4.95 14
9 19.90 4.46 25
10 23.30 3.81 12
――――――――――――――――――――――――――――――

Example 4 (+)-4,5-Dihydroxy-4- (trifluoromethyl) -3- {1- [5- (trifluoromethyl) pyrazin-2-yl] piperidin-4-yl} -1 , 4,5,7-Tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one hydrate (+)-4,5-dihydroxy-4-prepared in Example 1 (4) (Trifluoromethyl) -3- {1- [5- (trifluoromethyl) pyrazin-2-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4 b] Water (393 μl) was added to pyridin-6-one (19.64 mg) at room temperature. Thereafter, the mixture was stirred at 40 ° C. for about 20 hours, and then stirred at room temperature for about 0.5 hour. The solid was collected by filtration and dried overnight at room temperature to obtain the title compound (14.93 mg). 71% recovery.

Calculated as elemental analysis value C ₁₇ H ₁₆ F ₆ N ₆ O ₃ · 1.75H ₂ O: C, 41.01; H, 3.95; F, 22.90; N, 16.88.
Found: C, 40.95; H, 3.81; F, 23.24; N, 16.66.

Table 4 shows the diffraction pattern of powder X-ray diffraction (CuKα, λ = 1.54 Å, scanning speed = 20 ° / min), and the peak having a relative intensity of 11 or more when the maximum peak intensity is 100 in FIG.

(Table 4)
――――――――――――――――――――――――――――――
Peak number 2θ d value Relative intensity ――――――――――――――――――――――――――――――
1 9.52 9.28 94
2 12.14 7.28 20
3 14.36 6.16 11
4 14.86 5.96 42
5 17.76 4.99 18
6 18.24 4.86 17
7 18.74 4.73 100
8 19.56 4.53 14
9 25.60 3.48 14
10 29.32 3.04 11
――――――――――――――――――――――――――――――

Example 5 (+)-4,5-Dihydroxy-4- (trifluoromethyl) -3- {1- [5- (trifluoromethyl) pyrazin-2-yl] piperidin-4-yl} -1 , 4,5,7-Tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one (+)-4,5-dihydroxy-4- (trifluoro) prepared in Example 1 (4) Methyl) -3- {1- [5- (trifluoromethyl) pyrazin-2-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridine After -6-one (20.40 mg) was heat-treated at 250 ° C. with a thermal analyzer, toluene (408 μl) was added at room temperature. Thereafter, the mixture was stirred at 10 ° C. for about 20 hours, and then stirred at room temperature for about 0.5 hour. The solid was collected by filtration and dried overnight at room temperature to obtain the title compound (13.53 mg). Recovery 66%.

Calculated as Elemental Analysis Value C ₁₇ H ₁₆ F ₆ N ₆ O ₃ : C, 43.78; H, 3.46; F, 24.44; N, 18.02.
Found: C, 43.58; H, 3.41; F, 24.17; N, 17.78.

Table 5 shows the diffraction pattern of powder X-ray diffraction (CuKα, λ = 1.54 Å, scanning speed = 20 ° / min) and the peak with a relative intensity of 17 or more when the maximum peak intensity is 100 in FIG.

(Table 5)
――――――――――――――――――――――――――――――
Peak number 2θ d value Relative intensity ――――――――――――――――――――――――――――――
1 4.70 18.79 17
2 12.82 6.90 25
3 14.26 6.21 26
4 15.60 5.68 26
5 15.94 5.56 34
6 18.50 4.79 100
7 19.44 4.56 55
8 19.98 4.44 20
9 23.66 3.76 26
10 25.90 3.44 44
――――――――――――――――――――――――――――――

Example 6 (+)-cis-5-hydroxy-4- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl} -1 , 4,5,7-Tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one hydrate (1) 4- [1- (diphenylmethyl) -6-oxo-4- (trifluoromethyl) ) -4,5,6,7-tetrahydro-1H-pyrazolo [3,4-b] pyridin-3-yl] piperidine-1-carboxylate tert-butyl

Tert-Butyl 4- [5-amino-1- (diphenylmethyl) -1H-pyrazol-3-yl] piperidine-1-carboxylate (2.50 g, 5.78 mmol) prepared in Example 1 (1) ) And Meldrum's acid (2.50 g, 17.3 mmol) in ethanol (50 mL) were added trifluoroacetaldehyde ethyl hemiacetal (2.78 g, 17.3 mmol), and the mixture was stirred with heating under reflux for 3 hours. The solvent of the reaction solution was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography [NH-silica gel, elution solvent: hexane / ethyl acetate = 95 / 5-50 / 50 (gradient)]. Fractions containing the target compound were collected and triturated with ethyl acetate and hexane to obtain the title compound (1.48 g, yield: 46%).

¹ H-NMR (400Hz, CDCl ₃ ) δ: 7.44-7.12 (10H, m), 6.69 (1H, s), 4.13 (1H, brs), 3.65-3.55 (1H, m), 2.95-2.69 (7H, m), 1.87-1.55 (4H, m), 1.45 (9H, s).

(2) 3- [1- (tert-Butoxycarbonyl) piperidin-4-yl] -1- (diphenylmethyl) -5-hydroxy-6-oxo-4- (trifluoromethyl) -4,5,6 Methyl 7-tetrahydro-1H-pyrazolo [3,4-b] pyridine-5-carboxylate

4- [1- (Diphenylmethyl) -6-oxo-4- (trifluoromethyl) -4,5,6,7-tetrahydro-1H-pyrazolo [3,4-b] prepared in (1) To a solution of tert-butyl pyridin-3-yl] piperidine-1-carboxylate (2.92 g, 5.27 mmol) and dimethyl carbonate (0.665 mL, 7.90 mmol) in THF (50 mL) at −78 ° C. Lithium diisopropylamide (hexane-THF solution, 14.5 mL, 15.8 mmol) was added dropwise, the cooling bath was removed, and the mixture was stirred for 30 minutes while naturally heating. A saturated aqueous ammonium chloride solution was added to the reaction solution, followed by extraction three times with ethyl acetate. The obtained organic layer was dried over magnesium sulfate, and the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel chromatography [NH-silica gel, elution solvent: dichloromethane / methanol = 100 / 0-90 / 10 (gradient)] to obtain a synthetic intermediate.

1,8-diazabicyclo [5.4.0] -7-undecene (hereinafter referred to as DBU, 1.57 mL, 10.5 mmol), to a THF (50 mL) solution of the synthetic intermediate obtained by the above operation, (1S)-(+)-(10-camphorsulfonyl) oxaziridine (0.725 g, 3.16 mmol) and (1R)-(−)-(10-camphorsulfonyl) oxaziridine (0.725 g, 3.16 mmol) ) And stirred at room temperature for 2 hours. Saturated aqueous ammonium chloride solution was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The resulting organic layer was washed successively with saturated aqueous sodium bicarbonate solution and saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure. Distilled off. The obtained residue was purified by silica gel column chromatography [eluent: dichloromethane / methanol = 99 / 1-90 / 10 (gradient)] to give the title compound (2.77 g, yield: 84%). .

¹ H-NMR (400MHz, DMSO-d ₆ ) δ: 11.52 (1H, s), 7.38-7.17 (10H, m), 6.86 (1H, s), 4.12-3.89 (3H, m), 3.73 (3H, s), 2.89-2.67 (3H, m), 1.81-1.29 (4H, m), 1.39 (9H, s).

(3) (+)-4- [cis-1- (diphenylmethyl) -5-hydroxy-6-oxo-4- (trifluoromethyl) -4,5,6,7-tetrahydro-1H-pyrazolo [3 , 4-b] pyridin-3-yl] piperidine-1-carboxylate tert-butyl

3- [1- (tert-Butoxycarbonyl) piperidin-4-yl] -1- (diphenylmethyl) -5-hydroxy-6-oxo-4- (trifluoromethyl) -4 prepared in (2) , 5,6,7-tetrahydro-1H-pyrazolo [3,4-b] pyridine-5-carboxylate (4.36 g, 6.94 mmol) 1,4-dioxane (50 mL) and water (20 mL ) Lithium hydroxide monohydrate (0.873 g, 20.8 mmol) was added to the mixed solution, and the mixture was stirred at 50 ° C. for 1 hour. The reaction mixture was cooled to room temperature, saturated aqueous ammonium chloride solution was added, and the mixture was extracted with ethyl acetate. The obtained organic layer was washed successively with saturated aqueous sodium bicarbonate solution and saturated brine, dried over anhydrous sodium sulfate, and dried under reduced pressure. The solvent was distilled off. The obtained residue was purified by silica gel column chromatography [eluent: hexane / ethyl acetate = 95 / 5-50 / 50 (gradient)] to obtain a synthetic intermediate.

A part of the synthetic intermediate (1.23 g) obtained by the above operation was dissolved in ethyl acetate, neutral silica gel was added and adsorbed, and the solvent was distilled off under reduced pressure. The obtained powder was purified by flash LC [column: Chiralflash IC (30 mm id × 100 mm); manufactured by Daicel, elution solvent: hexane / ethanol = 91/9, flow rate: 12 mL / min] to give the title compound ( 0.55 g, yield: 27%, optically active substance).

Optical purity was measured using HPLC [column: Chiralpak IA (4.6 mm id x 250 mm); manufactured by Daicel, elution solvent: hexane / IPA = 70/30, flow rate 1.0 mL / min].

Optical purity 99% or more (retention time: 4.3 minutes);
¹ H-NMR (400MHz, DMSO-d ₆ ) δ: 11.21 (1H, s), 7.37-7.15 (10H, m), 6.74 (1H, s), 5.79 (1H, d, J = 4Hz), 4.57- 4.54 (1H, m), 4.16-3.90 (3H, m), 2.86-2.39 (3H, m), 1.83-1.35 (4H, m), 1.39 (9H, s);
[α] _D ²⁵ = + 35 ° (DMF, c = 1.00).

(4) (+)-cis-5-hydroxy-4- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl} -1,4 , 5,7-Tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one

(+)-4- [cis-1- (diphenylmethyl) -5-hydroxy-6-oxo-4- (trifluoromethyl) -4,5,6,7-tetrahydro- produced in (3) To a mixed solution of tert-butyl 1H-pyrazolo [3,4-b] pyridin-3-yl] piperidine-1-carboxylate (0.52 g, 0.91 mmol) in dichloromethane (25 mL) and acetonitrile (10 mL), Chlorotrimethylsilane (0.31 mL, 2.5 mmol) and sodium iodide (0.31 g, 2.1 mmol) were added, and the mixture was stirred at room temperature for 2 hours. The solvent of the reaction solution was distilled off under reduced pressure, a saturated aqueous sodium hydrogen carbonate solution and ethyl acetate were added to separate the solution, and the resulting organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and dried under reduced pressure. The solvent was distilled off.

To a solution of the obtained residue in DMSO (30 mL), 5-chloro-2- (trifluoromethyl) pyrimidine (0.37 mL, 2.0 mmol) and DBU (0.62 mL, 4.2 mmol) were added. Stir at 18 ° C. for 18 hours. The reaction mixture was diluted with ethyl acetate, washed successively with water and saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel column chromatography [eluent: hexane / ethyl acetate = 95 / 5-50 / 50 (gradient)] to obtain a synthetic intermediate.

Triethylsilane (0.200 mL, 1.26 mmol) and trifluoroacetic acid (1.0 mL, 13 mmol) were added to a solution of the synthetic intermediate obtained in the above operation in dichloromethane (3 mL), and the mixture was added at room temperature for 1 hour. Stir. The solvent of the reaction solution was distilled off, and ethyl acetate and saturated aqueous sodium hydrogen carbonate solution were added for liquid separation, and the obtained organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and reduced pressure. The solvent was distilled off. The obtained residue was purified by silica gel column chromatography [elution solvent: ethyl acetate / methanol = 50 / 50-0 / 100 (gradient)] to give the title compound (0.11 g, yield: 26%, Optically active substance) was obtained.

¹ H-NMR (400MHz, DMSO-d ₆ ) δ: 12.22 (1H, s), 10.55 (1H, s), 8.67 (2H, s), 5.53 (1H, d, J = 3Hz), 4.44 (1H, d, J = 6Hz), 4.25-4.10 (3H, m), 3.14-2.95 (3H, m), 1.93-1.38 (4H, m);
MS (ESI) m / z: 451 (M + H) ⁺ ;
[α] _D ²⁵ = + 7.2 ° (DMF, c = 1.00).

(5) (+)-cis-5-hydroxy-4- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl} -1,4 , 5,7-Tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one hydrate (+)-cis-5-hydroxy-4- (trifluoromethyl) prepared in (4) -3- {1- [2- (trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridine-6 -On (19.93 mg) was added water (399 μl) at room temperature, then stirred at 40 ° C. for about 20 hours and then at room temperature for about 0.5 hour. The solid was collected by filtration and dried overnight at room temperature to obtain the title compound (17.63 mg). Recovery rate 85%.

Calculated as elemental analysis value C ₁₇ H ₁₆ F ₆ N ₆ O ₂ .1.0H ₂ O: C, 43.60; H, 3.87; F, 24.34; N, 17.94.
Found: C, 43.64; H, 3.87; F, 24.35; N, 17.68.

Table 6 shows the peak of relative intensity 20 or more when the diffraction pattern of powder X-ray diffraction (CuKα, λ = 1.54 Å, scanning speed = 20 ° / min) in FIG.

(Table 6)
――――――――――――――――――――――――――――――
Peak number 2θ d value Relative intensity ――――――――――――――――――――――――――――――
1 9.72 9.09 71
2 11.14 7.94 34
3 11.66 7.58 22
4 16.68 5.31 54
5 17.84 4.97 39
6 18.94 4.68 29
7 19.26 4.60 68
8 19.70 4.50 100
9 23.00 3.86 20
10 25.42 3.50 94
――――――――――――――――――――――――――――――

Example 7 (+)-cis-5-hydroxy-4- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl} -1 , 4,5,7-Tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one hydrate (+)-cis-5-hydroxy-4-prepared in Example 6 (4) (Trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4 b] Toluene (407 μl) was added to pyridin-6-one (20.37 mg) at room temperature, followed by stirring at 40 ° C. for about 20 hours, and then at room temperature for about 0.5 hour. The solid was collected by filtration and dried overnight at room temperature to obtain the title compound (18.10 mg). Recovery rate 85%.

Calculated as elemental analysis value C ₁₇ H ₁₆ F ₆ N ₆ O ₂ .1.0H ₂ O · 0.05C ₇ H ₈ : C, 44.06; H, 3.92; F, 24.10; N, 17.77.
Found: C, 44.27; H, 3.67; F, 24.37; N, 17.54.

Table 7 shows the diffraction pattern of powder X-ray diffraction (CuKα, λ = 1.54 angstrom, scanning speed = 20 ° / min) and the relative intensity of 58 or more when the maximum peak intensity is 100 in FIG.

(Table 7)
――――――――――――――――――――――――――――――
Peak number 2θ d value Relative intensity ――――――――――――――――――――――――――――――
1 3.68 23.99 86
2 14.86 5.96 58
3 16.86 5.25 59
4 17.06 5.19 64
5 17.48 5.07 68
6 17.94 4.94 90
7 18.94 4.68 67
8 19.36 4.58 77
9 19.62 4.52 62
10 20.18 4.40 100
――――――――――――――――――――――――――――――

Example 8 (+)-cis-5-hydroxy-4- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl} -1 , 4,5,7-Tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one (+)-cis-5-hydroxy-4- (trifluoro) prepared in Example 6 (4) Methyl) -3- {1- [2- (trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridine Toluene (402 μl) was added to −6-one (20.08 mg) at room temperature, and the mixture was stirred at 60 ° C. for about 20 hours, and then stirred at room temperature for about 0.5 hour. The solid was collected by filtration and dried overnight at room temperature to obtain the title compound (16.13 mg). Recovery rate 80%.

Calculated as Elemental Analysis Value C ₁₇ H ₁₆ F ₆ N ₆ O ₂ : C, 45.34; H, 3.58; F, 25.31; N, 18.66.
Found: C, 45.45; H, 3.61; F, 25.27; N, 18.28.

Table 8 shows the peak of relative intensity 4 or more when the diffraction pattern of powder X-ray diffraction (CuKα, λ = 1.54 Å, scanning speed = 20 ° / min) is 100 in FIG.

(Table 8)
――――――――――――――――――――――――――――――
Peak number 2θ d value Relative intensity ――――――――――――――――――――――――――――――
1 5.66 15.60 100
2 9.54 9.26 4
3 12.38 7.14 12
4 17.00 5.21 69
5 21.82 4.07 15
6 22.90 3.88 4
7 26.12 3.41 4
8 27.50 3.24 10
9 28.72 3.11 4
10 33.10 2.70 9
――――――――――――――――――――――――――――――

Example 9 (+)-cis-5-hydroxy-4- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl} -1 , 4,5,7-Tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one (+)-cis-5-hydroxy-4- (trifluoro) prepared in Example 6 (4) Methyl) -3- {1- [2- (trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridine After -6-one (20.58 mg) was heat-treated with a thermal analyzer to 280 ° C., toluene (412 μl) was added at room temperature. Thereafter, the mixture was stirred at 10 ° C. for about 20 hours, and then stirred at room temperature for about 0.5 hour. The solid was collected by filtration and dried overnight at room temperature to obtain the title compound (17.95 mg). Recovery rate 87%.

Calculated as Elemental Analysis Value C ₁₇ H ₁₆ F ₆ N ₆ O ₂ : C, 45.34; H, 3.58; F, 25.31; N, 18.66.
Found: C, 45.18; H, 3.50; F, 25.46; N, 18.38.

9 shows the diffraction pattern of powder X-ray diffraction (CuKα, λ = 1.54 Å, scanning speed = 20 ° / min), and peaks having a relative intensity of 30 or more when the maximum peak intensity is 100 in FIG.

(Table 9)
――――――――――――――――――――――――――――――
Peak number 2θ d value Relative intensity ――――――――――――――――――――――――――――――
1 8.94 9.88 72
2 14.48 6.11 62
3 15.86 5.58 74
4 17.42 5.09 30
5 17.70 5.01 34
6 18.00 4.92 93
7 20.38 4.35 100
8 23.20 3.83 38
9 23.64 3.76 57
10 27.16 3.28 31
――――――――――――――――――――――――――――――

Example 10 (+)-cis-5-hydroxy-4- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl} -1 , 4,5,7-Tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one (+)-cis-5-hydroxy-4- (trifluoro) prepared in Example 6 (4) Methyl) -3- {1- [2- (trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridine Nitromethane (571 μl) was added to a sample (57.12 mg) obtained by heating -6-one to 280 ° C. with a thermal analyzer at room temperature. Thereafter, the mixture was stirred at 10 ° C. for about 4 hours, and then stirred at room temperature for about 1.5 hours. The solid was collected by filtration and dried overnight at room temperature to obtain the title compound (27.81 mg). Recovery rate 49%.

Elemental analysis _{C 17 H 16 F 6 N 6} O 2 · 0.75H 2 O · 0.05C 7 H 3 NO 2 Calculated: C, 43.86; H, 3.81 ; F, 24.41; N, 18.15.
Found: C, 43.59; H, 3.57; F, 24.65; N, 18.28.

Table 10 shows a peak having a relative intensity of 23 or more when the diffraction peak pattern of powder X-ray diffraction (CuKα, λ = 1.54 Å, scanning speed = 20 ° / min) in FIG.

(Table 10)
――――――――――――――――――――――――――――――
Peak number 2θ d value Relative intensity ――――――――――――――――――――――――――――――
1 10.66 8.29 26
2 14.96 5.92 83
3 16.06 5.51 51
4 16.70 5.30 38
5 18.64 4.76 27
6 19.70 4.50 100
7 20.84 4.26 48
8 22.30 3.98 23
9 23.34 3.81 23
10 24.28 3.66 38
――――――――――――――――――――――――――――――

Example 11 (+)-cis-5R-hydroxy-4R- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl} -1 , 4,5,7-Tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one (+)-cis-5-hydroxy-4- (trifluoro) prepared in Example 6 (4) Methyl) -3- {1- [2- (trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridine Acetone (200 μl) was added to -6-one (20.07 mg) and completely dissolved. Thereafter, acetone was distilled off with a centrifugal concentrator, and n-butyl acetate (200 μl) was added to the residue. Thereafter, the mixture was stirred at 40 ° C. for about 4 hours and at room temperature for about 0.5 hour. The precipitated solid was collected by filtration and dried overnight at room temperature to obtain the title compound (14.43 mg). Recovery rate 72%.

Calculated as Elemental Analysis Value C ₁₇ H ₁₆ F ₆ N ₆ O ₂ : C, 45.34; H, 3.58; F, 25.31; N, 18.66.
Found: C, 45.27; H, 3.46; F, 25.47; N, 18.40.

Table 11 shows the peak of relative intensity 11 or more when the diffraction peak pattern of powder X-ray diffraction (CuKα, λ = 1.54 Å, scanning speed = 20 ° / min) is 100 in FIG.

(Table 11)
――――――――――――――――――――――――――――――
Peak number 2θ d value Relative intensity ――――――――――――――――――――――――――――――
1 7.36 12.00 18
2 10.36 8.53 31
3 14.52 6.10 15
4 16.34 5.42 100
5 19.46 4.56 46
6 20.52 4.32 11
7 23.20 3.83 59
8 24.70 3.60 29
9 26.24 3.39 13
10 29.38 3.04 15
――――――――――――――――――――――――――――――

Example 12 (+)-cis-5R-hydroxy-4R- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl} -1 , 4,5,7-Tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one hydrate (+)-cis-5-hydroxy-4-prepared in Example 6 (4) (Trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidin-5-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4 b] 60% aqueous ethanol (200 μl) was added to pyridin-6-one (20.23 mg), and the mixture was stirred at 25 ° C. for about 24 hours and at room temperature for about 0.5 hour. The precipitated solid was collected by filtration and dried overnight at room temperature to obtain the title compound (19.20 mg). Recovery rate 82%.

Elemental analysis _{C 17 H 16 F 6 N 6} O 2 · 4.2H 2 O Calculated: C, 38.82; H, 4.68 ; F, 21.67; N, 15.98.
Found: C, 38.62; H, 4.56; F, 21.90; N, 15.82.

Table 12 shows the diffraction pattern of powder X-ray diffraction (CuKα, λ = 1.54 Å, scanning speed = 20 ° / min), and the peak having a relative intensity of 28 or more when the maximum peak intensity is 100 in FIG.

(Table 12)
――――――――――――――――――――――――――――――
Peak number 2θ d value Relative intensity ――――――――――――――――――――――――――――――
1 8.32 10.62 100
2 9.90 8.93 91
3 10.70 8.26 45
4 16.66 5.32 29
5 16.68 5.25 28
6 17.88 4.96 35
7 19.36 4.58 43
8 19.62 4.52 62
9 19.96 4.44 79
10 23.16 3.84 30
――――――――――――――――――――――――――――――

(Test Example 1) Measurement of LCAT activity (in vitro)
A density gradient centrifugation was performed to obtain a fraction composed of HDL3 (1.125 <specific gravity <1.210 g / mL) from plasma of a healthy person. The obtained fraction was dialyzed with phosphate buffered saline (pH 7.4) and used as an enzyme source and acceptor of LCAT. The test drug was prepared by dissolving in dimethyl sulfoxide. Phosphate buffered saline (pH 7.4) containing 1 mg / mL HDL3, DTNB (Ielman reagent, final concentration 0.5 mM), mercaptoethanol (final concentration 12.5 mM), and 0.6% bovine serum albumin [ ¹⁴ C] cholesterol containing, and various concentrations of test drug were added to make the total volume 80 μL. After this mixture was incubated at 37 ° C. for about 16 hours, a mixture of hexane and isopropanol (mixing ratio = 3: 2) was added to stop the reaction. After stirring, the hexane layer was collected and concentrated to dryness. Chloroform solution (concentration 10 mg / mL) was added thereto, spotted on a silica gel thin layer plate, and developed with a mixed solution of hexane, diethyl ether and ethyl acetate (mixing ratio = 85: 15: 2). The radioactivity of the portion corresponding to cholesterol oleate was measured with an imaging analyzer BAS-2500 (manufactured by Fuji Film). Samples to which no test drug was added were similarly processed and measured. Using the following equation, compared with no addition of test drug was calculated EC ₅₀ values of LCAT activation. The results are shown in Table 13.

In the formula, X represents the logarithm of the concentration of the test drug,
Y represents the response of the test drug (LCAT activity),
Top indicates the maximum value (maximum plateau),
“Bottom” indicates a minimum value (minimum flat area);
EC ₅₀ indicates a 50% effective concentration.

(Table 13)
――――――――――――――――――――――――――――――
Test compound EC ₅₀ (μM)
――――――――――――――――――――――――――――――
Compound of Example 1 (4) 0.004
Compound of Example 6 (4) 0.018
――――――――――――――――――――――――――――――

As described above, the compound of the present invention has an excellent LCAT activation action and is useful as a medicament for the treatment or prevention of diseases such as dyslipidemia and arteriosclerosis.

(Test Example 2) Measurement of LCAT activity (plasma)
Human, cynomolgus monkey or human LCAT transgenic mouse plasma is used as an enzyme source and acceptor for LCAT. The test drug is prepared by dissolving in dimethyl sulfoxide. [14C] cholesterol containing DTNB (Ielman's reagent, final concentration 0.5 mM), mercaptoethanol (final concentration 12.5 mM), and 0.6% bovine serum albumin was added to 5 μL of each plasma and 45 μL of PBS. Add the test drug at a concentration to make the total volume 80 μL. After incubating this mixture at 37 ° C. for about 16 hours, the reaction is stopped by adding a mixture of hexane and isopropanol (mixing ratio = 3: 2). After adding water and stirring, the hexane layer is collected and concentrated to dryness. Chloroform solution (concentration 10 mg / mL) is added to this, spotted on a silica gel thin layer plate, and developed with a mixed solution of hexane, diethyl ether and ethyl acetate (mixing ratio = 85: 15: 2). The radioactivity of the portion corresponding to cholesterol oleate is measured with an imaging analyzer BAS-2500 (manufactured by Fuji Film). The same processing and measurement is performed for the sample to which no test drug is added. Using the following formula, the EC ₅₀ value of LCAT activation is calculated in comparison with the case where no test drug is added.

In the formula, X represents the logarithm of the concentration of the test drug,
Y represents the response of the test drug (LCAT activity),
Top indicates the maximum value (maximum plateau),
“Bottom” indicates a minimum value (minimum flat area);
EC ₅₀ indicates a 50% effective concentration.

(Test Example 3) Measurement of LCAT activity (Ex vivo)
LCAT activity in plasma of cynomolgus monkeys or human LCAT transgenic mice administered with the test drug is measured. [25C] Each plasma is added with [14C] cholesterol containing DTNB (Ielman's reagent, final concentration 0.26 mM), mercaptoethanol (final concentration 2 mM), and 0.6% bovine serum albumin to a total volume of 40 μL. This mixture is incubated at 37 ° C. for 1 hour, and then the reaction is stopped by adding a mixture of hexane and isopropanol (mixing ratio = 3: 2). After adding water and stirring, the hexane layer is collected and concentrated to dryness. Chloroform solution (concentration 10 mg / mL) is added to this, spotted on a silica gel thin layer plate, and developed with a mixed solution of hexane, diethyl ether and ethyl acetate (mixing ratio = 85: 15: 2). The radioactivity of the portion corresponding to cholesterol oleate is measured with an imaging analyzer BAS-2500 (manufactured by Fuji Film). Compared with the LCAT activity before administration, the rate of change in LCAT activation at each time point is calculated.

(Test Example 4) Cynomolgus monkey efficacy test The test drug was propylene glycol (Sigma-Aldrich) -Tween 80 (Sigma-Aldrich) mixed solution [4/1 (v / v)] or 0.5% (w / v) It was dissolved in an aqueous methylcellulose solution and orally administered to cynomolgus monkeys for 1 or 7 days. Blood was collected before and after administration on the first or seventh day of administration to obtain plasma. Plasma cholesterol content was measured using a commercially available measurement kit (cholesterol-E Wako, Wako Pure Chemical Industries). The lipoprotein profile was analyzed by HPLC (column: Lipopropak XL, manufactured by Tosoh Corporation). The contents of HDL cholesterol and non-HDL cholesterol were calculated according to the following calculation formula.

HDL cholesterol content = plasma cholesterol content × (HDL cholesterol peak area / sum of each peak)
non-HDL cholesterol content = plasma cholesterol content × (peak area of non-HDL cholesterol / sum of each peak)
The increase rate (%) of HDL after a single administration of 10 mg / kg as compared to before administration was determined from AUC before administration and 24 hours after administration. The results are shown in Table 14.

(Table 14)
――――――――――――――――――――――――――――――
Test compound Increased HDL after a single dose -------------
Compound 590 of Example 1 (4)
Compound 483 of Example 6 (4)
――――――――――――――――――――――――――――――

(Test Example 5) Human LCAT transgenic mouse drug efficacy test The test drug was dissolved in polypropylene glycol-Tween 80 mixed solution [4/1 (v / v)] or 0.5% (w / v) methylcellulose aqueous solution, Human LCAT transgenic mice are orally administered for 1, 4 or 7 days. Blood is collected before and after administration on the first, fourth or seventh day of administration to obtain plasma. The cholesterol content in plasma is measured using a commercially available measurement kit (cholesterol-E Wako, Wako Pure Chemical Industries). The lipoprotein profile is analyzed by HPLC (column: Lipopropak XL, Tosoh). The content of HDL cholesterol and non-HDL cholesterol is calculated according to the following calculation formula.

HDL cholesterol content = plasma cholesterol content × (HDL cholesterol peak area / sum of each peak)
non-HDL cholesterol content = plasma cholesterol content × (peak area of non-HDL cholesterol / sum of each peak)
As described above, the compound of the present invention exhibits an excellent LCAT activation action and is useful as a medicament for the treatment or prevention of diseases such as dyslipidemia and arteriosclerosis.

Formulation Example 1 Hard Capsule Each standard bipartite hard gelatin capsule contains 100 mg of the powdered compound of Example 1, 150 mg lactose, 50 mg cellulose and 6 mg magnesium stearate. The unit capsule is manufactured by filling, and after washing, dried.

Formulation Example 2 Soft Capsules A mixture of the compound of Example 2 in a digestible oil such as soybean oil, cottonseed oil or olive oil is prepared and injected into gelatin with a positive displacement pump, Soft capsules containing 100 mg of active ingredient are obtained, washed and dried.

Formulation Example 3 Tablet According to conventional methods, 100 mg of the compound of Example 3, 0.2 mg colloidal silicon dioxide, 5 mg magnesium stearate, 275 mg microcrystalline cellulose, 11 mg starch and 98.8 mg Manufactured using lactose.

In addition, if desired, apply a coating.

(Formulation example 4) Suspension agent In 5 mL, 100 mg of the compound of Example 4 finely divided, 100 mg sodium carboxymethyl cellulose, 5 mg sodium benzoate, 1.0 g sorbitol solution (Japanese Pharmacopoeia ) And 0.025 mL of vanillin.

(Formulation Example 5) Injection: 1.5% by weight of the compound of Example 6 is stirred in 10% by weight of propylene glycol, adjusted to a certain volume with water for injection, and then sterilized to give an injection. .

The compound represented by the general formula (I) of the present invention or a pharmacologically acceptable salt thereof has an excellent LCAT activation action, and in particular, arteriosclerosis, arteriosclerotic heart disease, coronary heart disease. (Including acute coronary syndrome, heart failure, myocardial infarction, angina, cardiac ischemia, cardiovascular disorders and angiogenic restenosis), cerebrovascular disease (including stroke and cerebral infarction), peripheral vascular disease (peripheral arterial disease, Treatment of diabetic vascular complications), dyslipidemia, LCAT deficiency, low HDL cholesterolemia, high LDL cholesterolemia, diabetes, hypertension, metabolic syndrome, Alzheimer's disease, corneal opacity, or renal disease or It is useful as an active ingredient of prophylactic agents, particularly anti-arteriosclerotic agents.

Claims (48)

1. Formula (I)
   

   

   
   [Wherein, R ¹ represents a hydrogen atom or a hydroxyl group, and R represents a 2- (trifluoromethyl) pyrimidin-5-yl group or a 5- (trifluoromethyl) pyrazin-2-yl group. Or a pharmacologically acceptable salt thereof.
2. The crystal according to claim 1, wherein R ¹ is a hydrogen atom, and R is a 2- (trifluoromethyl) pyrimidin-5-yl group.
3. The crystal according to claim 1, wherein R ¹ is a hydroxyl group, and R is a 5- (trifluoromethyl) pyrazin-2-yl group.
4. (+)-4,5-dihydroxy-4- (trifluoromethyl) -3- {1- [5- (trifluoromethyl) pyrazin-2-yl] piperidin-4-yl} -1,4,5 7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one or (+)-cis-5-hydroxy-4- (trifluoromethyl) -3- {1- [2- (tri Fluoromethyl) pyrimidin-5-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one, or a hydrate crystal thereof.
5. In the powder X-ray diffraction pattern obtained by irradiation of copper with Kα rays, the interplanar spacing d is 8.86, 7.19, 6.67, 6.07, 5.51, 4.76, 4.61, 3. (+)-4,5-dihydroxy-4- (trifluoromethyl) -3- {1- [5- (trifluoromethyl) pyrazine-, showing major peaks at 94, 3.79 and 3.47 angstroms 2-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one hydrate crystals.
6. In the powder X-ray diffraction pattern obtained by irradiation with copper Kα rays, the interplanar spacing d is 15.28, 7.70, 6.67, 6.17, 5.62, 5.01, 4.58, 4. (+)-4,5-dihydroxy-4- (trifluoromethyl) -3- {1- [5- (trifluoromethyl) pyrazine-, showing major peaks at 43, 3.77 and 3.30 angstroms 2-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one hydrate crystals.
7. In the powder X-ray diffraction pattern obtained by irradiation of copper with Kα rays, the spacing d is 14.97, 11.50, 9.84, 9.50, 7.12, 5.73, 5.48, 4. (+)-4,5-dihydroxy-4- (trifluoromethyl) -3- {1- [5- (trifluoromethyl) pyrazine-, showing major peaks at 95, 4.46 and 3.81 Å 2-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one hydrate crystals.
8. In the powder X-ray diffraction pattern obtained by irradiation of copper with Kα rays, the interplanar spacing d is 9.28, 7.28, 6.16, 5.96, 4.99, 4.86, 4.73, 4. (+)-4,5-dihydroxy-4- (trifluoromethyl) -3- {1- [5- (trifluoromethyl) pyrazine-, showing major peaks at 53, 3.48 and 3.04 angstroms 2-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one hydrate crystals.
9. In the powder X-ray diffraction pattern obtained by irradiation with copper Kα rays, the interplanar spacing d is 18.79, 6.90, 6.21, 5.68, 5.56, 4.79, 4.56, 4. (+)-4,5-dihydroxy-4- (trifluoromethyl) -3- {1- [5- (trifluoromethyl) pyrazine-, showing major peaks at 44, 3.76 and 3.44 angstroms Crystals of 2-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one.
10. In the powder X-ray diffraction pattern obtained by irradiation of copper with Kα rays, the interplanar spacing d is 9.09, 7.94, 7.58, 5.31, 4.97, 4.68, 4.60, 4. (+)-Cis-5-hydroxy-4- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidine-, showing major peaks at 50, 3.86 and 3.50 angstroms 5-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one hydrate crystals.
11. In the powder X-ray diffraction pattern obtained by irradiation with copper Kα rays, the interplanar spacing d is 23.99, 5.96, 5.25, 5.19, 5.07, 4.94, 4.68, 4. (+)-Cis-5-hydroxy-4- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidine-, which exhibits major peaks at 58, 4.52 and 4.40 angstroms 5-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one hydrate crystals.
12. In the powder X-ray diffraction pattern obtained by irradiation with copper Kα rays, the interplanar spacing d is 15.60, 9.26, 7.14, 5.21, 4.07, 3.88, 3.41 and 3.41. (+)-Cis-5-hydroxy-4- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidine-, which exhibits major peaks at 24, 3.11, and 2.70 angstroms Crystals of 5-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one.
13. In the powder X-ray diffraction pattern obtained by irradiation with copper Kα rays, the interplanar spacing d is 9.88, 6.11, 5.58, 5.09, 5.01, 4.92, 4.35, 3. (+)-Cis-5-hydroxy-4- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidine-, showing major peaks at 83, 3.76 and 3.28 angstroms Crystals of 5-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one.
14. In the powder X-ray diffraction pattern obtained by irradiation of copper with Kα rays, the interplanar spacing d is 8.29, 5.92, 5.51, 5.30, 4.76, 4.50, 4.26, 3. (+)-Cis-5-hydroxy-4- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidine-, which shows major peaks at 98, 3.81 and 3.66 angstroms Crystals of 5-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one.
15. In the powder X-ray diffraction pattern obtained by irradiation with copper Kα rays, the interplanar spacing d is 12.00, 8.53, 6.10, 5.42, 4.56, 4.32, 3.83, 3. (+)-Cis-5R-hydroxy-4R- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidine-, which exhibits major peaks at 60, 3.39 and 3.04 angstroms Crystals of 5-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one.
16. In the powder X-ray diffraction pattern obtained by irradiation of copper with Kα rays, the interplanar spacing d is 10.62, 8.93, 8.26, 5.32, 5.25, 4.96, 4.58, 4. (+)-Cis-5R-hydroxy-4R- (trifluoromethyl) -3- {1- [2- (trifluoromethyl) pyrimidine-, which exhibits major peaks at 52, 4.44 and 3.84 angstroms 5-yl] piperidin-4-yl} -1,4,5,7-tetrahydro-6H-pyrazolo [3,4-b] pyridin-6-one hydrate crystals.
17. A pharmaceutical composition comprising the crystal according to any one of claims 1 to 16 as an active ingredient.
18. 18. The treatment for or prevention of arteriosclerosis, arteriosclerotic heart disease, coronary heart disease, cerebrovascular disease, peripheral vascular disease, dyslipidemia, low HDL cholesterolemia, or renal disease. Pharmaceutical composition.
19. The pharmaceutical composition according to claim 17 for the treatment or prevention of arteriosclerosis.
20. The pharmaceutical composition according to claim 17 for treating or preventing dyslipidemia.
21. The pharmaceutical composition according to claim 17, for the treatment or prevention of a disease caused by an increase in the concentration of LDL cholesterol in the blood.
22. 18. The pharmaceutical composition according to claim 17, for the treatment or prevention of a disease caused by a decrease in the concentration of HDL cholesterol in the blood.
23. An LCAT activator containing the crystal according to any one of claims 1 to 16 as an active ingredient.
24. A reversible LCAT activator comprising the crystal according to any one of claims 1 to 16 as an active ingredient.
25. An anti-arteriosclerotic agent comprising the crystal according to any one of claims 1 to 16 as an active ingredient.
26. A prophylactic or therapeutic agent for arteriosclerosis comprising the crystal according to any one of claims 1 to 16 as an active ingredient.
27. An agent for reducing the concentration of LDL cholesterol in blood, comprising the crystal according to any one of claims 1 to 16 as an active ingredient.
28. An agent for increasing the concentration of HDL cholesterol in blood, comprising the crystal according to any one of claims 1 to 16 as an active ingredient.
29. A pharmaceutical composition comprising the crystal according to any one of claims 1 to 16 and a pharmacologically acceptable carrier.
30. Use of the crystal according to any one of claims 1 to 16 for the production of a pharmaceutical composition.
31. Manufacture of a pharmaceutical composition for the treatment or prevention of arteriosclerosis, arteriosclerotic heart disease, coronary heart disease, cerebrovascular disease, peripheral vascular disease, dyslipidemia, low HDL cholesterolemia, or renal disease Use according to claim 30, for.
32. Use according to claim 30, for the manufacture of a pharmaceutical composition for the treatment or prevention of arteriosclerosis.
33. Use according to claim 30, for the manufacture of a pharmaceutical composition for the treatment or prevention of dyslipidemia.
34. Use according to claim 30, for the manufacture of a pharmaceutical composition for the treatment or prevention of diseases caused by an increase in the concentration of LDL cholesterol in the blood.
35. Use according to claim 30, for the manufacture of a pharmaceutical composition for the treatment or prevention of diseases caused by a decrease in the concentration of HDL cholesterol in the blood.
36. A method for activating LCAT, comprising administering an effective amount of the crystal according to any one of claims 1 to 16 to a human.
37. A method for treating or preventing a disease, comprising administering to a human an effective amount of the crystal according to any one of claims 1 to 16.
38. The disease is described in claim 37, wherein the disease is arteriosclerosis, arteriosclerotic heart disease, coronary heart disease, cerebrovascular disease, peripheral vascular disease, dyslipidemia, low HDL cholesterolemia, or renal disease. Method.
39. The method according to claim 37, wherein the disease is arteriosclerosis.
40. The method according to claim 37, wherein the disease is dyslipidemia.
41. The method according to claim 37, wherein the disease is a disease caused by an increase in the concentration of LDL cholesterol in the blood.
42. The method according to claim 37, wherein the disease is a disease caused by a decrease in the concentration of HDL cholesterol in the blood.
43. The crystal according to any one of claims 1 to 16, for use in a method for treatment or prevention of a disease.
44. 44. The disease according to claim 43, wherein the disease is arteriosclerosis, arteriosclerotic heart disease, coronary heart disease, cerebrovascular disease, peripheral vascular disease, dyslipidemia, low HDL cholesterolemia, or renal disease. Crystal.
45. 44. The crystal according to claim 43, wherein the disease is arteriosclerosis.
46. 44. The crystal according to claim 43, wherein the disease is dyslipidemia.
47. 44. The crystal according to claim 43, wherein the disease is a disease caused by an increase in the concentration of LDL cholesterol in the blood.
48. 44. The crystal of claim 43, wherein the disease is a disease caused by a decrease in the concentration of HDL cholesterol in the blood.
    
    

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